Widespread exposure to capsaicin occurs through food and topical medicines. To investigate potential food-drug or drug-drug interactions, capsaicin was evaluated in vitro against seven human drug-metabolizing cytochrome P450 (CYP) enzymes. At concentrations occurring after ingestion of chili peppers or topical administration of a high-concentration patch, capsaicin did not cause direct inhibition of any CYP enzyme. Direct inhibition was only observed at much higher concentrations; the lowest IC(50) value was 2.0 μM. For CYP2E1, the IC(50) value was too high to calculate. With pre-incubation, inhibition decreased for CYP1A2, 2C9, 2C19 and 3A4/5, whereas inhibition of CYP2B6 increased and moderately increased for CYP2D6. Induction of CYP activity was evaluated in microsomes from hepatocyte primary cultures. Capsaicin did not induce CYP1A2, 2B6, 2C9, 2C19, 2E1 or 3A4/5. 10 μM capsaicin caused a statistically significant increase in CYP1A2 activity (8.6% of the positive control). Inhibition of drug metabolism by capsaicin should be minimal, as the ratio of [I]/K(i) for direct inhibition is < 0.1. Although pre-incubation did enhance the potency for CYP2B6 inhibition to 5.1 μM, given that exposure to capsaicin from either food or a topical medicine is very low (≤58 nM) and transient, effects on CYPs appear unlikely.